Manual / Product Insert

Axiom™ 2.0 Assay Mini 96-Array Format Manual Protocol

Version: Rev. 1

Product FAQ

Which magnetic rack should I use to follow the manual protocol of MagJET kits?

Answer

Thermo Scientific offers MagJET Separation Rack of different formats for 1.5 mL tubes (Cat. Nos. MR01, MR02), 50 mL tubes (Cat. No. MR04), and 96-well plates (Cat. No. MR03). You may use other compatible magnetic racks available in the market as well.

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Manual / Product Insert

Manual: Protocol for Manual Synthesis of a Peptide (English )

Version: 20 Jun 2007

Product FAQ

After I added the chromogen reagent to the plate, I incubated the ELISA as suggested in the manual, but the A450 of the highest standard was higher than what my plate reader can read. What should I do?

Answer

Our customers use a wide variety of plate readers. Some of these can’t read absorbances higher than 2 AUFS (Absorbance Units Full Scale), while others can’t go beyond 3 AUFS, for example. If you read your ELISA plates after 30 minutes of incubation at room temperature and 1 or 2 A450 values are off-scale, you can shorten the incubation time. For example, some customers find that 20 minutes is the ideal incubation time because the ambient temperature in their lab is higher than approximately 2 degrees F (22 degrees C). In this case, higher temperatures increase the rate of the HRP-driven ELISA. Conversely, if the A450 values you get are not high enough, you can increase the incubation time accordingly.

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Product FAQ

I tried the antibody dilution (concentration) that you suggested in your manual, and it didn’t work. What should I do now?

Answer

The definition of “didn’t work” depends on what you did with the antibody. For example, it could mean that you observed no specific immunofluorescence staining at the antibody concentration that we suggested. It could also mean that, at the antibody dilution we recommended, your western blots showed high background staining. The suggested antibody concentrations specified in our manuals should be considered starting points for further experimentation. These values were derived in our labs during development of the antibody, or they are based on the experiences of our collaborators as well as other customers. If our suggestions don’t work for you, try your method again with a different concentration/dilution. Continue optimizing until your results are as good as you can make them. If your experiments still don’t “work”, please contact Technical Support at techsupport@thermofisher.com for further help.

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Product FAQ

Where can I find the protocols for my Ion Torrent Ion OneTouch 2 kit?

Answer

Ion Torrent OneTouch2 protocols can be found at our NGS Support Center (thermofisher.com/ngssupport), in the NGS Template Preparation Support Center section. For all other protocols, please go here (http://www.thermofisher.com/manuals-protocols-search.html).

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Product FAQ

Can I use a mini-prep kit for propagation of my lentiviral construct?

Answer

We do not recommend using a mini-prep kit for propagation of lentiviral constructs because the DNA yield from lentiviral mini-prep DNA is often very low due to the presence of the LTRs in the vector backbone. We recommend preparing lentiviral plasmid DNA using the S.N.A.P. MidiPrep Kit (Cat. No. K191001) or PureLink HiPure Plasmid Midiprep Kit (Cat. No. K210004), both of which contain 10 mM EDTA in the Resuspension Buffer. Since lentiviral DNA midi-preps also often have low DNA yields, we recommend following specific protocols to increase yield-basically, grow cells slowly, use fewer cells per column, and use 100 mL lentivirus culture for each DNA midi-prep.

Note: If you are going to be mini-prepping the lentiviral plasmid during the cloning/colony screening processes, we recommend using the PureLink HQ Mini Kit (Cat. No. K210001) and following the manual protocol with one change: only a single elution with 50 mL TE, pH 8.0 buffer. The typical yield with this method is normally pretty low: 100-150 ng/mL (i.e., 5-7 mg total). The OD 260/280 is typically between 1.8 and 2.1.

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Product FAQ

I used a mini-prep kit for propagating my lentiviral construct and obtained very low DNA yield. Can you please offer some tips?

Answer

The DNA yield from lentiviral mini-prep DNA is often very low due to the presence of the LTRs in the vector backbone. Hence, we do not recommend using a mini-prep kit for propagation of lentiviral constructs. We recommend preparing lentiviral plasmid DNA using the S.N.A.P. MidiPrep Kit (Cat. No. K191001) or the PureLink HiPure Plasmid Midiprep Kit (Cat. No. K210004), both of which contain 10 mM EDTA in the Resuspension Buffer. Since lentiviral DNA midi-preps also often have low DNA yields, we recommend following specific protocols to increase yield- basically, grow cells slowly, use fewer cells per column, and use 100 mL lentivirus culture for each DNA midi-prep.

Note: If you are going to be mini-prepping the lentiviral plasmid during the cloning/colony screening processes, we recommend using the PureLink HQ Mini Kit (Cat. No. K210001) and following the manual protocol with one change: only a single elution with 50 mL TE, pH 8.0 buffer. The typical yield with this method is normally pretty low, 100-150 ng/mL (i.e., 5-7 mg total). The OD 260/280 is typically between 1.8 and 2.1.

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